This invention relates to optical sectioning microscopy.
Sectioning microscopy can be performed using structured illumination, which may also be referred to as patterned excitation.
Wide-field optical microscopy is one technique for observing biological specimens. For a traditional wide-field microscope, the resolving power (e.g., the ability of its optical components to distinguish two spatially separate points of a specimen) is constrained by the diffraction limit of light. According to the Rayleigh criterion, two points of a specimen can be resolved if they are separated at least by ˜0.5λ laterally and ˜λ in depth (λ being the wavelength of the light). This resolution is insufficient for imaging many small objects such as sub-cellular organelles.
In the past decades, various resolution-enhancing techniques have been developed for achieving a sub-diffraction-limit optical resolution (sometimes referred to as “super-resolution”) in wide-field microscopes. Despite their success, many of these techniques may not be suited for implementations in ordinary laboratories, for example, due to system complexity and/or high costs. Some computational methods, on the other hand, apply numerically iterative image processing algorithms for resolving high-contrast images (such as fluorescence or topography images). Still, the computation time for processing time-lapse movies can be overly long when dynamic analyses of a specimen are desired.